Jump to navigation Jump to search
Protocol: -Glycerol Stock:
- Add 1 culture from selection plate into 7.5 mL of LB Broth with 50 μg/mL of KAN
- Incubated culture overnight at 37°C with 225 rpm
- Started at 5pm 22/04/2019.
- Stopped at 8:30 am 23/04/2019
- Mix 0.5 mL of overnight culture with 0.5 mL of 50% glycerol in LB broth.
- Store glycerol stocks in -80°C freezer
- Use the remaining 5 mL of the overnight culture (mentioned in the glycerol stock protocol)
- Inoculate 200 mL of LB Broth with 50 μg/mL of KAN with the 5 mL overnight culture.
- Incubated culture for 2 hours at 37°C with 225 rpm.
- Add 205 μL of 1 M IPTG to the culture.
- Continue to incubate the culture at 37°C with 225 rpm for 4.5 hours.
- Split culture into four 50 mL falcon tubes and centrifuge at 3750 rpm for 30 mins.
- Remove supernatant and store pellet in -20°C freezer.
Results: The pellets after expression were all very pink. Protein production appears to be successful.